ESTB_BURGA
ID ESTB_BURGA Reviewed; 392 AA.
AC Q9KX40;
DT 05-MAY-2009, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2000, sequence version 1.
DT 25-MAY-2022, entry version 78.
DE RecName: Full=Esterase EstB;
DE EC=3.1.1.-;
GN Name=estB;
OS Burkholderia gladioli (Pseudomonas marginata) (Phytomonas marginata).
OC Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales;
OC Burkholderiaceae; Burkholderia.
OX NCBI_TaxID=28095;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], CHARACTERIZATION AS AN ESTERASE,
RP MUTAGENESIS OF SER-75 AND SER-149, ABSENCE OF BETA-LACTAMASE ACTIVITY, AND
RP PROBABLE SUBCELLULAR LOCATION.
RC STRAIN=ATCC 10248 / NCPPB 1891;
RX PubMed=11472796; DOI=10.1016/s0168-1656(01)00284-x;
RA Petersen E.I., Valinger G., Soelkner B., Stubenrauch G., Schwab H.;
RT "A novel esterase from Burkholderia gladioli which shows high deacetylation
RT activity on cephalosporins is related to beta-lactamases and DD-
RT peptidases.";
RL J. Biotechnol. 89:11-25(2001).
RN [2]
RP RANDOM MUTAGENESIS TO CREATE MORE STABLE ORGANIC SOLVENT-RESISTANT ENZYME.
RC STRAIN=ATCC 10248 / NCPPB 1891;
RX PubMed=17137667; DOI=10.1016/j.jbiotec.2006.10.008;
RA Valinger G., Hermann M., Wagner U.G., Schwab H.;
RT "Stability and activity improvement of cephalosporin esterase EstB from
RT Burkholderia gladioli by directed evolution and structural interpretation
RT of muteins.";
RL J. Biotechnol. 129:98-108(2007).
RN [3]
RP RANDOM MUTAGENESIS TO ALTER ENANTIOSELECTIVITY.
RC STRAIN=ATCC 10248 / NCPPB 1891;
RX PubMed=17147964; DOI=10.1016/j.jbiotec.2006.10.007;
RA Ivancic M., Valinger G., Gruber K., Schwab H.;
RT "Inverting enantioselectivity of Burkholderia gladioli esterase EstB by
RT directed and designed evolution.";
RL J. Biotechnol. 129:109-122(2007).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS) WITH AND WITHOUT THE INHIBITOR
RP DIISOPROPYL-FLUOROPHOSPHATE.
RC STRAIN=ATCC 10248 / NCPPB 1891;
RX PubMed=11847270; DOI=10.1110/ps.33002;
RA Wagner U.G., Petersen E.I., Schwab H., Kratky C.;
RT "EstB from Burkholderia gladioli: a novel esterase with a beta-lactamase
RT fold reveals steric factors to discriminate between esterolytic and beta-
RT lactam cleaving activity.";
RL Protein Sci. 11:467-478(2002).
CC -!- FUNCTION: Acts on short-chain (C4-C6) fatty acid esters and
CC triglycerides, including tertiary alcohol esters. Activity on p-
CC nitrophenyl esters is generally higher than on o-nitrophenyl esters.
CC Lacks beta-lactamase activity; it hydrolyzes the ester bond of
CC cephalosporin substrates but there is no opening of the beta-lactam
CC ring observed.
CC -!- ACTIVITY REGULATION: Strongly inhibited by eserin, NaF, HgCl2, SDS and
CC Triton X-100.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.3 mM for cephalosporin C;
CC KM=1.0 mM for 7-amino cephalosporinic acid;
CC Vmax=79 umol/min/mg enzyme for cephalosporin C;
CC Vmax=77 umol/min/mg enzyme for 7-amino cephalosporinic acid;
CC Note=Esterase, not beta-lactamase activity of the enzyme.;
CC pH dependence:
CC Optimum pH is 7.0.;
CC Temperature dependence:
CC Optimum temperature is 43 degrees Celsius.;
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- BIOTECHNOLOGY: Directed evolutionary mutagenesis has created an enzyme
CC with 40-fold higher stability in organic solvents. It contains the
CC following changes: Leu-8, Ser-132, Arg-134, Cys-155, Gly-251, Val-311
CC and Lys-316 and could have applications in antibiotic synthesis
CC (PubMed:17137667). {ECO:0000269|PubMed:17137667}.
CC -!- SIMILARITY: Belongs to the class-A beta-lactamase family.
CC {ECO:0000305}.
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DR EMBL; U33634; AAF59826.1; -; Genomic_DNA.
DR RefSeq; WP_036056391.1; NZ_PVHE01000058.1.
DR PDB; 1CI8; X-ray; 2.00 A; A/B=1-392.
DR PDB; 1CI9; X-ray; 1.80 A; A/B=1-392.
DR PDBsum; 1CI8; -.
DR PDBsum; 1CI9; -.
DR AlphaFoldDB; Q9KX40; -.
DR SMR; Q9KX40; -.
DR DrugBank; DB04491; Diisopropylphosphono Group.
DR MEROPS; S12.950; -.
DR KEGG; bgo:BM43_5119; -.
DR EvolutionaryTrace; Q9KX40; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0016787; F:hydrolase activity; IEA:UniProtKB-KW.
DR Gene3D; 3.40.710.10; -; 1.
DR InterPro; IPR001466; Beta-lactam-related.
DR InterPro; IPR012338; Beta-lactam/transpept-like.
DR InterPro; IPR023650; Beta-lactam_class-A_AS.
DR Pfam; PF00144; Beta-lactamase; 1.
DR SUPFAM; SSF56601; SSF56601; 1.
DR PROSITE; PS00146; BETA_LACTAMASE_A; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Hydrolase.
FT CHAIN 1..392
FT /note="Esterase EstB"
FT /id="PRO_0000370699"
FT ACT_SITE 75
FT /note="Acyl-ester intermediate"
FT MUTAGEN 75
FT /note="S->A: Complete loss of esterase activity."
FT /evidence="ECO:0000269|PubMed:11472796"
FT MUTAGEN 149
FT /note="S->A: Retains 30% esterase activity."
FT /evidence="ECO:0000269|PubMed:11472796"
FT HELIX 16..31
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 36..44
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 47..58
FT /evidence="ECO:0007829|PDB:1CI9"
FT TURN 59..62
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 70..72
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 74..76
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 77..90
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 100..102
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 121..125
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 134..137
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 142..146
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 151..153
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 159..168
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 184..196
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 200..207
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 209..212
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 224..226
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 235..237
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 245..248
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 255..259
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 261..265
FT /evidence="ECO:0007829|PDB:1CI9"
FT TURN 273..275
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 281..293
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 295..298
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 300..307
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 313..315
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 322..324
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 326..332
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 334..337
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 346..350
FT /evidence="ECO:0007829|PDB:1CI9"
FT TURN 351..353
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 354..359
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 360..362
FT /evidence="ECO:0007829|PDB:1CI9"
FT STRAND 364..373
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 375..378
FT /evidence="ECO:0007829|PDB:1CI9"
FT HELIX 380..389
FT /evidence="ECO:0007829|PDB:1CI9"
SQ SEQUENCE 392 AA; 41707 MW; 60EC4764C3C499B7 CRC64;
MTAASLDPTA FSLDAASLAA RLDAVFDQAL RERRLVGAVA IVARHGEILY RRAQGLADRE
AGRPMREDTL FRLASVTKPI VALAVLRLVA RGELALDAPV TRWLPEFRPR LADGSEPLVT
IHHLLTHTSG LGYWLLEGAG SVYDRLGISD GIDLRDFDLD ENLRRLASAP LSFAPGSGWQ
YSLALDVLGA VVERATGQPL AAAVDALVAQ PLGMRDCGFV SAEPERFAVP YHDGQPEPVR
MRDGIEVPLP EGHGAAVRFA PSRVFEPGAY PSGGAGMYGS ADDVLRALEA IRANPGFLPE
TLADAARRDQ AGVGAETRGP GWGFGYLSAV LDDPAAAGTP QHAGTLQWGG VYGHSWFVDR
ALGLSVLLLT NTAYEGMSGP LTIALRDAVY AR
